ID IPA9_SHIFL Reviewed; 545 AA. AC Q8VSC3; O85159; Q2TH74; Q6XVV1; Q9AFM5; Q9AJV1; DT 13-JUL-2010, integrated into UniProtKB/Swiss-Prot. DT 04-JAN-2005, sequence version 2. DT 27-MAR-2024, entry version 122. DE RecName: Full=E3 ubiquitin-protein ligase ipaH9.8 {ECO:0000305}; DE EC=2.3.2.27 {ECO:0000269|PubMed:18005683, ECO:0000269|PubMed:18997778, ECO:0000269|PubMed:31216343, ECO:0000269|PubMed:33303953}; DE AltName: Full=Invasion plasmid antigen 9.8 {ECO:0000303|PubMed:9582283}; GN Name=ipaH9.8 {ECO:0000303|PubMed:15950937}; OrderedLocusNames=CP0226; GN ORFNames=pWR501_0234, SFLP090; OS Shigella flexneri. OG Plasmid pCP301, Plasmid pWR100, Plasmid pWR501, Plasmid pINV_F6_M1382, and OG Plasmid pSF5. OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Enterobacteriaceae; Shigella. OX NCBI_TaxID=623; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-12 AND 480-494, RP AND INDUCTION. RC STRAIN=M90T / Serotype 5a; PLASMID=pWR100; RX PubMed=9582283; DOI=10.1093/emboj/17.10.2894; RA Demers B., Sansonetti P.J., Parsot C.; RT "Induction of type III secretion in Shigella flexneri is associated with RT differential control of transcription of genes encoding secreted RT proteins."; RL EMBO J. 17:2894-2903(1998). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-9, AND SUBCELLULAR RP LOCATION. RC STRAIN=M90T / Serotype 5a; PLASMID=pWR100; RX PubMed=11115111; DOI=10.1046/j.1365-2958.2000.02179.x; RA Buchrieser C., Glaser P., Rusniok C., Nedjari H., d'Hauteville H., RA Kunst F., Sansonetti P.J., Parsot C.; RT "The virulence plasmid pWR100 and the repertoire of proteins secreted by RT the type III secretion apparatus of Shigella flexneri."; RL Mol. Microbiol. 38:760-771(2000). RN [3] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=M90T / Serotype 5a; PLASMID=pWR501; RX PubMed=11292750; DOI=10.1128/iai.69.5.3271-3285.2001; RA Venkatesan M.M., Goldberg M.B., Rose D.J., Grotbeck E.J., Burland V., RA Blattner F.R.; RT "Complete DNA sequence and analysis of the large virulence plasmid of RT Shigella flexneri."; RL Infect. Immun. 69:3271-3285(2001). RN [4] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=M1382 / Serotype 6; PLASMID=pINV_F6_M1382; RX PubMed=14573649; DOI=10.1128/iai.71.11.6298-6306.2003; RA Lan R., Stevenson G., Reeves P.R.; RT "Comparison of two major forms of the Shigella virulence plasmid pINV: RT positive selection is a major force driving the divergence."; RL Infect. Immun. 71:6298-6306(2003). RN [5] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=Serotype 5; PLASMID=pSF5; RX PubMed=16704117; DOI=10.1007/s11427-006-0141-3; RA Xiong Z., Tang X., Yang F., Zhang X., Yang J., Chen L., Nie H., Yan Y., RA Jiang Y., Wang J., Xue Y., Xu X., Zhu Y., Dong J., An L., Wang X., Jin Q.; RT "Comparison of the virulence plasmid genomes of two strains of Shigella RT which lost the ability to bind Congo red."; RL Sci. China, Ser. C, Life Sci. 49:141-148(2006). RN [6] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=301 / Serotype 2a; PLASMID=pCP301; RX PubMed=12384590; DOI=10.1093/nar/gkf566; RA Jin Q., Yuan Z., Xu J., Wang Y., Shen Y., Lu W., Wang J., Liu H., Yang J., RA Yang F., Zhang X., Zhang J., Yang G., Wu H., Qu D., Dong J., Sun L., RA Xue Y., Zhao A., Gao Y., Zhu J., Kan B., Ding K., Chen S., Cheng H., RA Yao Z., He B., Chen R., Ma D., Qiang B., Wen Y., Hou Y., Yu J.; RT "Genome sequence of Shigella flexneri 2a: insights into pathogenicity RT through comparison with genomes of Escherichia coli K12 and O157."; RL Nucleic Acids Res. 30:4432-4441(2002). RN [7] RP SUBCELLULAR LOCATION, AND SECRETION VIA TYPE III SECRETION SYSTEM. RC STRAIN=YSH6000 / Serotype 2a; RX PubMed=11418613; DOI=10.1074/jbc.m101882200; RA Toyotome T., Suzuki T., Kuwae A., Nonaka T., Fukuda H., Imajoh-Ohmi S., RA Toyofuku T., Hori M., Sasakawa C.; RT "Shigella protein IpaH(9.8) is secreted from bacteria within mammalian RT cells and transported to the nucleus."; RL J. Biol. Chem. 276:32071-32079(2001). RN [8] RP FUNCTION, DISRUPTION PHENOTYPE, AND INTERACTION WITH MURINE U2AF1. RC STRAIN=YSH6000 / Serotype 2a; RX PubMed=15950937; DOI=10.1016/j.bbrc.2005.05.145; RA Okuda J., Toyotome T., Kataoka N., Ohno M., Abe H., Shimura Y., RA Seyedarabi A., Pickersgill R., Sasakawa C.; RT "Shigella effector IpaH9.8 binds to a splicing factor U2AF(35) to modulate RT host immune responses."; RL Biochem. Biophys. Res. Commun. 333:531-539(2005). RN [9] RP FUNCTION AS A LIGASE, PATHWAY, CATALYTIC ACTIVITY, AND MUTAGENESIS OF RP CYS-337. RX PubMed=18005683; DOI=10.1016/j.chom.2007.02.002; RA Rohde J.R., Breitkreutz A., Chenal A., Sansonetti P.J., Parsot C.; RT "Type III secretion effectors of the IpaH family are E3 ubiquitin RT ligases."; RL Cell Host Microbe 1:77-83(2007). RN [10] RP FUNCTION, PATHWAY, CATALYTIC ACTIVITY, AND MUTAGENESIS OF CYS-337; GLU-338; RP ASP-339; ARG-340; ARG-387 AND ASP-397. RX PubMed=18997778; DOI=10.1038/nsmb.1511; RA Singer A.U., Rohde J.R., Lam R., Skarina T., Kagan O., Dileo R., RA Chirgadze N.Y., Cuff M.E., Joachimiak A., Tyers M., Sansonetti P.J., RA Parsot C., Savchenko A.; RT "Structure of the Shigella T3SS effector IpaH defines a new class of E3 RT ubiquitin ligases."; RL Nat. Struct. Mol. Biol. 15:1293-1301(2008). RN [11] RP FUNCTION, AND MUTAGENESIS OF CYS-337. RC STRAIN=YSH6000 / Serotype 2a; RX PubMed=20010814; DOI=10.1038/ncb2006; RA Ashida H., Kim M., Schmidt-Supprian M., Ma A., Ogawa M., Sasakawa C.; RT "A bacterial E3 ubiquitin ligase IpaH9.8 targets NEMO/IKKgamma to dampen RT the host NF-kappaB-mediated inflammatory response."; RL Nat. Cell Biol. 12:66-73(2010). RN [12] RP MUTAGENESIS OF ARG-163; PHE-187; HIS-210 AND CYS-337, ACTIVE SITE, AND RP FUNCTION. RC STRAIN=M90T / Serotype 5a; PLASMID=pWR100; RX PubMed=24248594; DOI=10.1128/mcb.01360-13; RA Keszei A.F., Tang X., McCormick C., Zeqiraj E., Rohde J.R., Tyers M., RA Sicheri F.; RT "Structure of an SspH1-PKN1 complex reveals the basis for host substrate RT recognition and mechanism of activation for a bacterial E3 ubiquitin RT ligase."; RL Mol. Cell. Biol. 34:362-373(2014). RN [13] RP FUNCTION, AND CATALYTIC ACTIVITY. RX PubMed=29024643; DOI=10.1016/j.chom.2017.09.007; RA Wandel M.P., Pathe C., Werner E.I., Ellison C.J., Boyle K.B., RA von der Malsburg A., Rohde J., Randow F.; RT "GBPs inhibit motility of Shigella flexneri but are targeted for RT degradation by the bacterial ubiquitin ligase IpaH9.8."; RL Cell Host Microbe 22:507-518(2017). RN [14] RP FUNCTION, SUBCELLULAR LOCATION, CATALYTIC ACTIVITY, ACTIVE SITE, AND RP MUTAGENESIS OF LEU-50; ARG-62; ASN-83 AND CYS-337. RX PubMed=29144452; DOI=10.1038/nature24467; RA Li P., Jiang W., Yu Q., Liu W., Zhou P., Li J., Xu J., Xu B., Wang F., RA Shao F.; RT "Ubiquitination and degradation of GBPs by a Shigella effector to suppress RT host defence."; RL Nature 551:378-383(2017). RN [15] RP FUNCTION. RX PubMed=37014865; DOI=10.1073/pnas.2218469120; RA Goers L., Kim K., Stedman T.C., Canning P.J., Mou X., Ernst N.H., Coers J., RA Lesser C.F.; RT "Shigella IpaH9.8 limits GBP1-dependent LPS release from intracytosolic RT bacteria to suppress caspase-4 activation."; RL Proc. Natl. Acad. Sci. U.S.A. 120:e2218469120-e2218469120(2023). RN [16] RP X-RAY CRYSTALLOGRAPHY (3.20 ANGSTROMS) OF 254-545, FUNCTION, INTERACTION RP WITH HUMAN U2AF1, AUTOUBIQUITINATION, AND ACTIVITY REGULATION. RC STRAIN=YSH6000 / Serotype 2a; RX PubMed=20831869; DOI=10.1016/j.febslet.2010.09.006; RA Seyedarabi A., Sullivan J.A., Sasakawa C., Pickersgill R.W.; RT "A disulfide driven domain swap switches off the activity of Shigella RT IpaH9.8 E3 ligase."; RL FEBS Lett. 584:4163-4168(2010). RN [17] {ECO:0007744|PDB:5B0N, ECO:0007744|PDB:5B0T} RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF 22-244, AND DOMAIN. RX PubMed=27050259; DOI=10.1107/s2053230x16002715; RA Takagi K., Kim M., Sasakawa C., Mizushima T.; RT "Crystal structure of the substrate-recognition domain of the Shigella E3 RT ligase IpaH9.8."; RL Acta Crystallogr. F 72:269-275(2016). RN [18] {ECO:0007744|PDB:6K2D} RP X-RAY CRYSTALLOGRAPHY (3.60 ANGSTROMS) OF 22-252 IN COMPLEX WITH HOST GBP1, RP FUNCTION, PATHWAY, CATALYTIC ACTIVITY, ACTIVITY REGULATION, AND MUTAGENESIS RP OF 86-TYR--GLN-88; HIS-126; TYR-146; ARG-190 AND CYS-337. RX PubMed=31216343; DOI=10.1371/journal.ppat.1007876; RA Ji C., Du S., Li P., Zhu Q., Yang X., Long C., Yu J., Shao F., Xiao J.; RT "Structural mechanism for guanylate-binding proteins (GBPs) targeting by RT the Shigella E3 ligase IpaH9.8."; RL PLoS Pathog. 15:e1007876-e1007876(2019). RN [19] {ECO:0007744|PDB:6LOJ} RP X-RAY CRYSTALLOGRAPHY (3.72 ANGSTROMS) IN COMPLEX WITH HOST GBP1, FUNCTION, RP PATHWAY, CATALYTIC ACTIVITY, ACTIVITY REGULATION, DOMAIN, AND MUTAGENESIS RP OF ARG-166; ILE-196; GLU-198; ILE-211; LEU-216; HIS-219 AND PHE-395. RX PubMed=33303953; DOI=10.1038/s42003-020-01492-1; RA Ye Y., Xiong Y., Huang H.; RT "Substrate-binding destabilizes the hydrophobic cluster to relieve the RT autoinhibition of bacterial ubiquitin ligase IpaH9.8."; RL Commun. Biol. 3:752-752(2020). CC -!- FUNCTION: Effector E3 ubiquitin ligase that interferes with host's CC ubiquitination pathway and modulates the acute inflammatory responses, CC thus facilitating bacterial colonization within the host cell CC (PubMed:18005683, PubMed:18997778, PubMed:20831869, PubMed:24248594, CC PubMed:29024643, PubMed:29144452, PubMed:31216343, PubMed:33303953). CC Interacts with IKBKG (NEMO) and TNIP1 (ABIN-1), a ubiquitin-binding CC adapter protein, which results in TNIP1-dependent 'Lys-27'-linked CC polyubiquitination of IKBKG (PubMed:18005683). Consequently, CC polyubiquitinated IKBKG undergoes proteasome-dependent degradation, CC which perturbs NF-kappa-B activation during bacterial infection CC (PubMed:18005683, PubMed:20010814). Mediates polyubiquitination of host CC U2AF1, leading to its proteasomal degradation (PubMed:15950937). CC Catalyzes 'Lys-48'-linked polyubiquitination and subsequent degradation CC of a subset of host guanylate-binding proteins (GBP1, GBP2, GBP4 and CC GBP6), thereby suppressing host cell defense (PubMed:29024643, CC PubMed:29144452, PubMed:31216343, PubMed:33303953, PubMed:37014865). In CC contrast, host GBP3 and GBP7 are not ubiquitinated by IpaH9.8 CC (PubMed:29024643, PubMed:29144452, PubMed:31216343). Uses UBE2D2 CC (UBCH5B) as an E2 ubiquitin-conjugating enzyme (PubMed:18005683). CC {ECO:0000269|PubMed:15950937, ECO:0000269|PubMed:18005683, CC ECO:0000269|PubMed:18997778, ECO:0000269|PubMed:20010814, CC ECO:0000269|PubMed:20831869, ECO:0000269|PubMed:24248594, CC ECO:0000269|PubMed:29024643, ECO:0000269|PubMed:29144452, CC ECO:0000269|PubMed:31216343, ECO:0000269|PubMed:33303953, CC ECO:0000269|PubMed:37014865}. CC -!- CATALYTIC ACTIVITY: CC Reaction=S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine + CC [acceptor protein]-L-lysine = [E2 ubiquitin-conjugating enzyme]-L- CC cysteine + N(6)-ubiquitinyl-[acceptor protein]-L-lysine.; CC EC=2.3.2.27; Evidence={ECO:0000269|PubMed:18005683, CC ECO:0000269|PubMed:18997778, ECO:0000269|PubMed:31216343, CC ECO:0000269|PubMed:33303953}; CC -!- ACTIVITY REGULATION: Exists in an autoinhibited state in the absence of CC substrate protein, due to interactions of the leucine-rich repeats with CC NEL domain (PubMed:20831869, PubMed:31216343, PubMed:33303953). Is CC activated upon binding to a substrate protein (PubMed:20831869, CC PubMed:31216343, PubMed:33303953). {ECO:0000269|PubMed:20831869, CC ECO:0000269|PubMed:31216343, ECO:0000269|PubMed:33303953}. CC -!- PATHWAY: Protein modification; protein ubiquitination. CC {ECO:0000269|PubMed:18005683, ECO:0000269|PubMed:18997778, CC ECO:0000269|PubMed:31216343, ECO:0000269|PubMed:33303953}. CC -!- SUBUNIT: Interacts also with human and mouse U2AF1 (U2AF35). CC {ECO:0000269|PubMed:15950937, ECO:0000269|PubMed:20831869}. CC -!- INTERACTION: CC Q8VSC3; Q8VSC3: ipaH9.8; NbExp=3; IntAct=EBI-6125799, EBI-6125799; CC Q8VSC3; Q9Y6K9: IKBKG; Xeno; NbExp=8; IntAct=EBI-6125799, EBI-81279; CC Q8VSC3; Q15025: TNIP1; Xeno; NbExp=4; IntAct=EBI-6125799, EBI-357849; CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:11115111}. Host CC cytoplasm {ECO:0000269|PubMed:11418613, ECO:0000269|PubMed:29144452}. CC Host nucleus {ECO:0000269|PubMed:11418613}. Note=Secreted via Mxi-Spa CC type III secretion system (T3SS), and delivered into the host cytoplasm CC (PubMed:11115111, PubMed:11418613). Transported into the host nucleus CC (PubMed:11418613). This transport is independent of cytosolic factors, CC but dependent on temperature and partly on ATP/GTP (PubMed:11418613). CC {ECO:0000269|PubMed:11115111, ECO:0000269|PubMed:11418613}. CC -!- INDUCTION: Induced upon entry into host epithelial cells. CC {ECO:0000269|PubMed:9582283}. CC -!- DOMAIN: The LRR (leucine-rich repeat) repeats are involved in substrate CC recognition with target proteins. {ECO:0000269|PubMed:27050259, CC ECO:0000269|PubMed:31216343}. CC -!- PTM: Autoubiquitinated (in vitro) (PubMed:20831869). Ubiquitinated in CC the presence of host E1 ubiquitin-activating enzyme, E2 ubiquitin- CC conjugating enzyme and ubiquitin (PubMed:20831869). CC {ECO:0000269|PubMed:20831869}. CC -!- DISRUPTION PHENOTYPE: In murine lung infection model, mutants cause CC severe inflammatory responses, with increased pro-inflammatory cytokine CC production levels. Colonization is greatly reduced. CC {ECO:0000269|PubMed:15950937}. CC -!- SIMILARITY: Belongs to the LRR-containing bacterial E3 ligase family. CC {ECO:0000305}. CC -!- SEQUENCE CAUTION: CC Sequence=AAK18544.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305}; CC Sequence=AAW64849.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305}; CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AF047365; AAC23714.1; -; Genomic_DNA. DR EMBL; AL391753; CAC05853.1; -; Genomic_DNA. DR EMBL; AF348706; AAK18544.1; ALT_INIT; Genomic_DNA. DR EMBL; AY206445; AAP79029.1; -; Genomic_DNA. DR EMBL; AY879342; AAW64849.1; ALT_INIT; Genomic_DNA. DR EMBL; AF386526; AAL72345.2; -; Genomic_DNA. DR RefSeq; NP_858359.2; NC_004851.1. DR RefSeq; YP_006960318.1; NC_019197.1. DR PDB; 3L3P; X-ray; 3.20 A; A=254-545. DR PDB; 5B0N; X-ray; 1.80 A; A/B=22-244. DR PDB; 5B0T; X-ray; 2.00 A; A=22-244. DR PDB; 6K2D; X-ray; 3.60 A; B=22-252. DR PDB; 6LOJ; X-ray; 3.72 A; A=22-244. DR PDB; 6LOL; X-ray; 2.75 A; A=22-545. DR PDBsum; 3L3P; -. DR PDBsum; 5B0N; -. DR PDBsum; 5B0T; -. DR PDBsum; 6K2D; -. DR PDBsum; 6LOJ; -. DR PDBsum; 6LOL; -. DR AlphaFoldDB; Q8VSC3; -. DR SMR; Q8VSC3; -. DR DIP; DIP-46341N; -. DR IntAct; Q8VSC3; 3. DR MINT; Q8VSC3; -. DR PaxDb; 198214-CP0226; -. DR GeneID; 1238048; -. DR KEGG; sfl:CP0226; -. DR PATRIC; fig|198214.7.peg.5483; -. DR HOGENOM; CLU_018533_2_0_6; -. DR UniPathway; UPA00143; -. DR PRO; PR:Q8VSC3; -. DR Proteomes; UP000001006; Plasmid pCP301. DR GO; GO:0005576; C:extracellular region; IDA:UniProtKB. DR GO; GO:0044164; C:host cell cytosol; IDA:UniProtKB. DR GO; GO:0042025; C:host cell nucleus; IDA:UniProtKB. DR GO; GO:0042802; F:identical protein binding; IPI:IntAct. DR GO; GO:0061630; F:ubiquitin protein ligase activity; IDA:UniProtKB. DR GO; GO:0004842; F:ubiquitin-protein transferase activity; IDA:UniProtKB. DR GO; GO:0043161; P:proteasome-mediated ubiquitin-dependent protein catabolic process; IDA:GO_Central. DR GO; GO:0051865; P:protein autoubiquitination; IDA:CACAO. DR GO; GO:0044314; P:protein K27-linked ubiquitination; IDA:UniProtKB. DR GO; GO:0070936; P:protein K48-linked ubiquitination; IDA:UniProtKB. DR GO; GO:0052036; P:suppression by symbiont of host inflammatory response; IDA:GO_Central. DR GO; GO:0044414; P:suppression of host defenses by symbiont; IDA:UniProt. DR GO; GO:0052170; P:suppression of host innate immune response; IDA:UniProtKB. DR GO; GO:0085034; P:symbiont-mediated suppression of host canonical NF-kappaB cascade; IDA:GO_Central. DR GO; GO:0006511; P:ubiquitin-dependent protein catabolic process; IDA:UniProt. DR Gene3D; 1.20.58.90; -; 1. DR Gene3D; 3.80.10.10; Ribonuclease Inhibitor; 1. DR Gene3D; 1.20.58.360; Shigella T3SS effector IpaH defines; 1. DR Gene3D; 1.20.1270.130; Shigella T3SS effector IpaH domain; 1. DR InterPro; IPR032675; LRR_dom_sf. DR InterPro; IPR032674; LRR_E3_ligase_N. DR InterPro; IPR029487; NEL_dom. DR PANTHER; PTHR47114; -; 1. DR PANTHER; PTHR47114:SF2; OLIGODENDROCYTE-MYELIN GLYCOPROTEIN; 1. DR Pfam; PF14496; NEL; 1. DR Pfam; PF12468; TTSSLRR; 1. DR SMART; SM00364; LRR_BAC; 5. DR SUPFAM; SSF52058; L domain-like; 1. PE 1: Evidence at protein level; KW 3D-structure; Direct protein sequencing; Host cytoplasm; Host nucleus; KW Leucine-rich repeat; Plasmid; Reference proteome; Repeat; Secreted; KW Transferase; Ubl conjugation; Ubl conjugation pathway; Virulence. FT CHAIN 1..545 FT /note="E3 ubiquitin-protein ligase ipaH9.8" FT /id="PRO_0000395755" FT REPEAT 57..77 FT /note="LRR 1" FT REPEAT 78..99 FT /note="LRR 2" FT REPEAT 100..117 FT /note="LRR 3" FT REPEAT 118..139 FT /note="LRR 4" FT REPEAT 140..157 FT /note="LRR 5" FT REPEAT 158..179 FT /note="LRR 6" FT REPEAT 182..203 FT /note="LRR 7" FT REPEAT 205..228 FT /note="LRR 8" FT DOMAIN 261..471 FT /note="NEL" FT /evidence="ECO:0000255" FT REGION 1..242 FT /note="Interaction with target proteins" FT /evidence="ECO:0000250|UniProtKB:P0CE12" FT REGION 243..250 FT /note="Linker" FT /evidence="ECO:0000250|UniProtKB:P0CE12" FT REGION 251..545 FT /note="E3 ubiquitin-protein ligase catalytic domain" FT /evidence="ECO:0000250|UniProtKB:P0CE12" FT ACT_SITE 337 FT /note="Glycyl thioester intermediate" FT /evidence="ECO:0000269|PubMed:24248594, FT ECO:0000269|PubMed:29144452" FT SITE 166 FT /note="Sensor for substrate-binding" FT /evidence="ECO:0000269|PubMed:33303953" FT SITE 187 FT /note="Sensor for substrate-binding" FT /evidence="ECO:0000269|PubMed:33303953" FT VARIANT 222 FT /note="P -> Q (in plasmid pWR100, plasmid pWR501, plasmid FT pSF5 and plasmid pINV_F6_M1382)" FT VARIANT 474 FT /note="Q -> L (in plasmid pINV_F6_M1382)" FT VARIANT 536 FT /note="P -> F (in plasmid pWR100 and plasmid pWR501)" FT VARIANT 536 FT /note="P -> S (in plasmid pSF5)" FT MUTAGEN 50 FT /note="L->A: Abolished ability to bind and ubiquitinate FT host GBP1; when associated with A-62 and A-83." FT /evidence="ECO:0000269|PubMed:29144452" FT MUTAGEN 62 FT /note="R->A: Abolished ability to bind and ubiquitinate FT host GBP1; when associated with A-50 and A-83." FT /evidence="ECO:0000269|PubMed:29144452" FT MUTAGEN 83 FT /note="N->A: Abolished ability to bind and ubiquitinate FT host GBP1; when associated with A-50 and A-62." FT /evidence="ECO:0000269|PubMed:29144452" FT MUTAGEN 86..88 FT /note="YNQ->ANA: Decreased ability to ubiquitinate host FT GBP1." FT /evidence="ECO:0000269|PubMed:31216343" FT MUTAGEN 126 FT /note="H->A: Decreased ability to ubiquitinate host GBP1; FT when associated with A-146." FT /evidence="ECO:0000269|PubMed:31216343" FT MUTAGEN 146 FT /note="Y->A: Decreased ability to ubiquitinate host GBP1; FT when associated with A-126 or A-190." FT /evidence="ECO:0000269|PubMed:31216343" FT MUTAGEN 163 FT /note="R->A: Abolishes proteasomal degradation of host FT proteins; when associated with A-187 and A-210." FT /evidence="ECO:0000269|PubMed:24248594" FT MUTAGEN 166 FT /note="R->A: Strongly reduced ability to ubiquitinate host FT GBP1." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 187 FT /note="F->A: Abolishes proteasomal degradation of host FT proteins; when associated with A-163 and A-210." FT /evidence="ECO:0000269|PubMed:24248594" FT MUTAGEN 190 FT /note="R->A: Decreased ability to ubiquitinate host GBP1; FT when associated with A-146." FT /evidence="ECO:0000269|PubMed:31216343" FT MUTAGEN 196 FT /note="I->A: Does not affect autoinhibition in absence of FT substrate." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 196 FT /note="I->D: Reduced autoinhibition in absence of FT substrate." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 198 FT /note="E->A: Does not affect autoinhibition in absence of FT substrate." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 210 FT /note="H->A: Abolishes proteasomal degradation of host FT proteins; when associated with A-163 and A-187." FT /evidence="ECO:0000269|PubMed:24248594" FT MUTAGEN 211 FT /note="I->D: Slightly reduced autoinhibition in absence of FT substrate." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 216 FT /note="L->D: Slightly reduced autoinhibition in absence of FT substrate." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 219 FT /note="H->A: Does not affect autoinhibition in absence of FT substrate." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 337 FT /note="C->A: Abolishes E3 ubiquitin ligase activity and FT proteasomal degradation of host proteins." FT /evidence="ECO:0000269|PubMed:18005683, FT ECO:0000269|PubMed:18997778, ECO:0000269|PubMed:20010814, FT ECO:0000269|PubMed:24248594, ECO:0000269|PubMed:29144452, FT ECO:0000269|PubMed:31216343" FT MUTAGEN 338 FT /note="E->A: Does not affect E3 ubiquitin ligase activity." FT /evidence="ECO:0000269|PubMed:18997778" FT MUTAGEN 339 FT /note="D->A: Abolishes E3 ubiquitin ligase activity." FT /evidence="ECO:0000269|PubMed:18997778" FT MUTAGEN 340 FT /note="R->A: Reduced E3 ubiquitin ligase activity." FT /evidence="ECO:0000269|PubMed:18997778" FT MUTAGEN 387 FT /note="R->A: Does not affect E3 ubiquitin ligase activity." FT /evidence="ECO:0000269|PubMed:18997778" FT MUTAGEN 395 FT /note="F->A: Does not affect autoinhibition in absence of FT substrate." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 395 FT /note="F->R: Reduced autoinhibition in absence of FT substrate." FT /evidence="ECO:0000269|PubMed:33303953" FT MUTAGEN 397 FT /note="D->A: Abolishes E3 ubiquitin ligase activity." FT /evidence="ECO:0000269|PubMed:18997778" FT CONFLICT 7 FT /note="Missing (in Ref. 2; AA sequence)" FT /evidence="ECO:0000305" FT HELIX 23..35 FT /evidence="ECO:0007829|PDB:5B0N" FT HELIX 43..55 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 59..62 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 80..83 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 101..103 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 121..123 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 141..143 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 161..163 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 185..187 FT /evidence="ECO:0007829|PDB:5B0N" FT HELIX 198..201 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 208..210 FT /evidence="ECO:0007829|PDB:5B0N" FT HELIX 220..229 FT /evidence="ECO:0007829|PDB:5B0N" FT STRAND 237..239 FT /evidence="ECO:0007829|PDB:5B0N" FT HELIX 254..259 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 264..273 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 274..276 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 283..293 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 305..318 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 320..359 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 366..386 FT /evidence="ECO:0007829|PDB:3L3P" FT TURN 387..389 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 400..408 FT /evidence="ECO:0007829|PDB:3L3P" FT TURN 409..414 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 434..448 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 451..455 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 458..466 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 469..484 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 486..498 FT /evidence="ECO:0007829|PDB:3L3P" FT HELIX 504..530 FT /evidence="ECO:0007829|PDB:3L3P" SQ SEQUENCE 545 AA; 61979 MW; 5E70042D797B5099 CRC64; MLPINNNFSL PQNSFYNTIS GTYADYFSAW DKWEKQALPG EERDEAVSRL KECLINNSDE LRLDRLNLSS LPDNLPAQIT LLNVSYNQLT NLPELPVTLK KLYSASNKLS ELPVLPPALE SLQVQHNELE NLPALPDSLL TMNISYNEIV SLPSLPQALK NLRATRNFLT ELPAFSEGNN PVVREYFFDR NQISHIPESI LNLRNECSIH ISDNPLSSHA LPALQRLTSS PDYHGPRIYF SMSDGQQNTL HRPLADAVTA WFPENKQSDV SQIWHAFEHE EHANTFSAFL DRLSDTVSAR NTSGFREQVA AWLEKLSASA ELRQQSFAVA ADATESCEDR VALTWNNLRK TLLVHQASEG LFDNDTGALL SLGREMFRLE ILEDIARDKV RTLHFVDEIE VYLAFQTMLA EKLQLSTAVK EMRFYGVSGV TANDLRTAEA MVRSREENEF TDWFSLWGPW HAVLKRTEAD RWAQAEEQKY EMLENEYPQR VADRLKASGL SGDADAEREA GAQVMRETEQ QIYRQLTDEV LALRLPENGS QLHHS //